Accumulating evidence has suggested that adipose tissue play a key role in obesity-associated increases in systemic inflammatory responses. It has been shown that adipose tissue mass positively correlates with and increased circulating inflammatory cytokine interleukin-6 (IL-6) and n-3 polyunsaturated fatty acids (n-3 PUFA) reduce obesity-associated inflammation. Recently results from our laboratory showed that subset of adipose tissue cells including adipose stem cells secrete significantly higher levels of inflammatory factors that can be accounted for an increased systemic inflammation. However, in obese mice no studies have addressed whether n-3 PUFA has an inhibitory role in IL-6 secretion and IL-6 gene expression in adipose tissue derived stem cells. Thus, we treated adipose stem cells with stearidonic acid (SDA, 18:4 n-3, EPA precursor) and eicosapentaenoic acid (EPA, 20:5 n-3) to determine if IL-6 secretion and gene expression were altered. Results demonstrated that EPA and SDA significantly reduced lipopolysaccharide (LPS)-induced IL-6 secretion and IL-6 mRNA expression levels in the adipose tissue stem cells. Toll-like-receptor 2 (TLR2) mediates this inhibitory effect in reducing IL-6 mRNA by inhibiting NF-κB activation and not via MAPK activation pathways.